Comparative Analysis of Colon Cancer-Derived Fusobacterium nucleatum Subspecies: Inflammation and Colon Tumorigenesis in Murine Models
ABSTRACT Fusobacteria are commonly associated with human colorectal cancer (CRC), but investigations are hampered by the absence of a stably colonized murine model. Further, Fusobacterium nucleatum subspecies isolated from human CRC have not been investigated. While F. nucleatum subspecies are commo...
| Main Authors: | , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2022-02-01
|
| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.02991-21 |
| _version_ | 1819280517830279168 |
|---|---|
| author | Jessica Queen Jada C. Domingue James Robert White Courtney Stevens Barath Udayasuryan Tam T. D. Nguyen Shaoguang Wu Hua Ding Hongni Fan Madison McMann Alina Corona Tatianna C. Larman Scott S. Verbridge Franck Housseau Daniel J. Slade Julia L. Drewes Cynthia L. Sears |
| author_facet | Jessica Queen Jada C. Domingue James Robert White Courtney Stevens Barath Udayasuryan Tam T. D. Nguyen Shaoguang Wu Hua Ding Hongni Fan Madison McMann Alina Corona Tatianna C. Larman Scott S. Verbridge Franck Housseau Daniel J. Slade Julia L. Drewes Cynthia L. Sears |
| author_sort | Jessica Queen |
| collection | DOAJ |
| description | ABSTRACT Fusobacteria are commonly associated with human colorectal cancer (CRC), but investigations are hampered by the absence of a stably colonized murine model. Further, Fusobacterium nucleatum subspecies isolated from human CRC have not been investigated. While F. nucleatum subspecies are commonly associated with CRC, their ability to induce tumorigenesis and contributions to human CRC pathogenesis are uncertain. We sought to establish a stably colonized murine model and to understand the inflammatory potential and virulence genes of human CRC F. nucleatum, representing the 4 subspecies, animalis, nucleatum, polymorphum, and vincentii. Five human CRC-derived and two non-CRC derived F. nucleatum strains were tested for colonization, tumorigenesis, and cytokine induction in specific-pathogen-free (SPF) and/or germfree (GF) wild-type and ApcMin/+ mice, as well as in vitro assays and whole-genome sequencing (WGS). SPF wild-type and ApcMin/+ mice did not achieve stable colonization with F. nucleatum, whereas certain subspecies stably colonized some GF mice but without inducing colon tumorigenesis. F. nucleatum subspecies did not form in vivo biofilms or associate with the mucosa in mice. In vivo inflammation was inconsistent across subspecies, whereas F. nucleatum induced greater cytokine responses in a human colorectal cell line, HCT116. While F. nucleatum subspecies displayed genomic variability, no distinct virulence genes associated with human CRC strains were identified that could reliably distinguish these strains from non-CRC clinical isolates. We hypothesize that the lack of F. nucleatum-induced tumorigenesis in our model reflects differences in human and murine biology and/or a synergistic role for F. nucleatum in concert with other bacteria to promote carcinogenesis. IMPORTANCE Colon cancer is a leading cause of cancer morbidity and mortality, and it is hypothesized that dysbiosis in the gut microbiota contributes to colon tumorigenesis. Fusobacterium nucleatum, a member of the oropharyngeal microbiome, is enriched in a subset of human colon tumors. However, it is unclear whether this genetically varied species directly promotes tumor formation, modulates mucosal immune responses, or merely colonizes the tumor microenvironment. Mechanistic studies to address these questions have been stymied by the lack of an animal model that does not rely on daily orogastric gavage. Using multiple murine models, in vitro assays with a human colon cancer cell line, and whole-genome sequencing analysis, we investigated the proinflammatory and tumorigenic potential of several F. nucleatum clinical isolates. The significance of this research is development of a stable colonization model of F. nucleatum that does not require daily oral gavages in which we demonstrate that a diverse library of clinical isolates do not promote tumorigenesis. |
| first_indexed | 2024-12-24T00:45:04Z |
| format | Article |
| id | doaj.art-0a9d26e8a85f4e6d870c28b44bda70be |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-24T00:45:04Z |
| publishDate | 2022-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-0a9d26e8a85f4e6d870c28b44bda70be2022-12-21T17:23:49ZengAmerican Society for MicrobiologymBio2150-75112022-02-0113110.1128/mbio.02991-21Comparative Analysis of Colon Cancer-Derived Fusobacterium nucleatum Subspecies: Inflammation and Colon Tumorigenesis in Murine ModelsJessica Queen0Jada C. Domingue1James Robert White2Courtney Stevens3Barath Udayasuryan4Tam T. D. Nguyen5Shaoguang Wu6Hua Ding7Hongni Fan8Madison McMann9Alina Corona10Tatianna C. Larman11Scott S. Verbridge12Franck Housseau13Daniel J. Slade14Julia L. Drewes15Cynthia L. Sears16Department of Medicine, Johns Hopkins University, Baltimore, Maryland, USADepartment of Medicine, Johns Hopkins University, Baltimore, Maryland, USAResphera Biosciences, Baltimore, Maryland, USADepartment of Medicine, Johns Hopkins University, Baltimore, Maryland, USADepartment of Biomedical Engineering and Mechanics, Virginia Polytechnic Institute, Blacksburg, Virginia, USADepartment of Biochemistry, Virginia Polytechnic Institute, Blacksburg, Virginia, USADepartment of Medicine, Johns Hopkins University, Baltimore, Maryland, USADepartment of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USADepartment of Oncology, Johns Hopkins University, Baltimore, Maryland, USADepartment of Medicine, Johns Hopkins University, Baltimore, Maryland, USADepartment of Medicine, Johns Hopkins University, Baltimore, Maryland, USADivision of Gastrointestinal and Liver Pathology, Johns Hopkins University, Baltimore, Maryland, USADepartment of Biomedical Engineering and Mechanics, Virginia Polytechnic Institute, Blacksburg, Virginia, USADepartment of Oncology, Johns Hopkins University, Baltimore, Maryland, USADepartment of Biochemistry, Virginia Polytechnic Institute, Blacksburg, Virginia, USADepartment of Medicine, Johns Hopkins University, Baltimore, Maryland, USADepartment of Medicine, Johns Hopkins University, Baltimore, Maryland, USAABSTRACT Fusobacteria are commonly associated with human colorectal cancer (CRC), but investigations are hampered by the absence of a stably colonized murine model. Further, Fusobacterium nucleatum subspecies isolated from human CRC have not been investigated. While F. nucleatum subspecies are commonly associated with CRC, their ability to induce tumorigenesis and contributions to human CRC pathogenesis are uncertain. We sought to establish a stably colonized murine model and to understand the inflammatory potential and virulence genes of human CRC F. nucleatum, representing the 4 subspecies, animalis, nucleatum, polymorphum, and vincentii. Five human CRC-derived and two non-CRC derived F. nucleatum strains were tested for colonization, tumorigenesis, and cytokine induction in specific-pathogen-free (SPF) and/or germfree (GF) wild-type and ApcMin/+ mice, as well as in vitro assays and whole-genome sequencing (WGS). SPF wild-type and ApcMin/+ mice did not achieve stable colonization with F. nucleatum, whereas certain subspecies stably colonized some GF mice but without inducing colon tumorigenesis. F. nucleatum subspecies did not form in vivo biofilms or associate with the mucosa in mice. In vivo inflammation was inconsistent across subspecies, whereas F. nucleatum induced greater cytokine responses in a human colorectal cell line, HCT116. While F. nucleatum subspecies displayed genomic variability, no distinct virulence genes associated with human CRC strains were identified that could reliably distinguish these strains from non-CRC clinical isolates. We hypothesize that the lack of F. nucleatum-induced tumorigenesis in our model reflects differences in human and murine biology and/or a synergistic role for F. nucleatum in concert with other bacteria to promote carcinogenesis. IMPORTANCE Colon cancer is a leading cause of cancer morbidity and mortality, and it is hypothesized that dysbiosis in the gut microbiota contributes to colon tumorigenesis. Fusobacterium nucleatum, a member of the oropharyngeal microbiome, is enriched in a subset of human colon tumors. However, it is unclear whether this genetically varied species directly promotes tumor formation, modulates mucosal immune responses, or merely colonizes the tumor microenvironment. Mechanistic studies to address these questions have been stymied by the lack of an animal model that does not rely on daily orogastric gavage. Using multiple murine models, in vitro assays with a human colon cancer cell line, and whole-genome sequencing analysis, we investigated the proinflammatory and tumorigenic potential of several F. nucleatum clinical isolates. The significance of this research is development of a stable colonization model of F. nucleatum that does not require daily oral gavages in which we demonstrate that a diverse library of clinical isolates do not promote tumorigenesis.https://journals.asm.org/doi/10.1128/mbio.02991-21Fusobacterium subspeciescolorectal cancermouse modelsFusobacterium genome sequencesFusobacterium virulenceFusobacterium |
| spellingShingle | Jessica Queen Jada C. Domingue James Robert White Courtney Stevens Barath Udayasuryan Tam T. D. Nguyen Shaoguang Wu Hua Ding Hongni Fan Madison McMann Alina Corona Tatianna C. Larman Scott S. Verbridge Franck Housseau Daniel J. Slade Julia L. Drewes Cynthia L. Sears Comparative Analysis of Colon Cancer-Derived Fusobacterium nucleatum Subspecies: Inflammation and Colon Tumorigenesis in Murine Models mBio Fusobacterium subspecies colorectal cancer mouse models Fusobacterium genome sequences Fusobacterium virulence Fusobacterium |
| title | Comparative Analysis of Colon Cancer-Derived Fusobacterium nucleatum Subspecies: Inflammation and Colon Tumorigenesis in Murine Models |
| title_full | Comparative Analysis of Colon Cancer-Derived Fusobacterium nucleatum Subspecies: Inflammation and Colon Tumorigenesis in Murine Models |
| title_fullStr | Comparative Analysis of Colon Cancer-Derived Fusobacterium nucleatum Subspecies: Inflammation and Colon Tumorigenesis in Murine Models |
| title_full_unstemmed | Comparative Analysis of Colon Cancer-Derived Fusobacterium nucleatum Subspecies: Inflammation and Colon Tumorigenesis in Murine Models |
| title_short | Comparative Analysis of Colon Cancer-Derived Fusobacterium nucleatum Subspecies: Inflammation and Colon Tumorigenesis in Murine Models |
| title_sort | comparative analysis of colon cancer derived fusobacterium nucleatum subspecies inflammation and colon tumorigenesis in murine models |
| topic | Fusobacterium subspecies colorectal cancer mouse models Fusobacterium genome sequences Fusobacterium virulence Fusobacterium |
| url | https://journals.asm.org/doi/10.1128/mbio.02991-21 |
| work_keys_str_mv | AT jessicaqueen comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT jadacdomingue comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT jamesrobertwhite comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT courtneystevens comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT barathudayasuryan comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT tamtdnguyen comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT shaoguangwu comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT huading comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT hongnifan comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT madisonmcmann comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT alinacorona comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT tatiannaclarman comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT scottsverbridge comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT franckhousseau comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT danieljslade comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT julialdrewes comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels AT cynthialsears comparativeanalysisofcoloncancerderivedfusobacteriumnucleatumsubspeciesinflammationandcolontumorigenesisinmurinemodels |